In Situ Synthesis and Self-Assembly of Peptide-PEG Conjugates: A Facile Method for the Construction of Fibrous Hydrogels.

Peptide-based hydrogels have gained considerable attention as a compelling platform for various biomedical applications in recent years. Their attractiveness stems from their ability to seamlessly integrate diverse properties, such as biocompatibility, biodegradability, easily adjustable hydrophilicity/hydrophobicity, and other functionalities. However, a significant drawback is that most of the functional self-assembling peptides cannot form robust hydrogels suitable for biological applications. In this study, we present the synthesis of novel peptide-PEG conjugates and explore their comprehensive hydrogel properties. The hydrogel comprises double networks, with the first network formed through the self-assembly of peptides to create a ß-sheet secondary structure. The second network is established through covalent bond formation via N-hydroxysuccinimide chemistry between peptides and a 4-arm PEG to form a covalently linked network. Importantly, our findings reveal that this hydrogel formation method can be applied to other peptides containing lysine-rich sequences. Upon encapsulation of the hydrogel with antimicrobial peptides, the hydrogel retained high bacterial killing efficiency while showing minimum cytotoxicity toward mammalian cells. We hope that this method opens new avenues for the development of a novel class of peptide-polymer hydrogel materials with enhanced performance in biomedical contexts, particularly in reducing the potential for infection in applications of tissue regeneration and drug delivery.

Supra-Alveolar Bone Regeneration: Progress, Challenges, and Future Perspectives.

Periodontitis is a highly prevalent disease that damages the supporting tissues of a tooth, including the alveolar bone. Alveolar bone loss owing to periodontitis is broadly categorized as supra-alveolar and intra-alveolar bone loss. In intra-alveolar bone loss, the defect has an angular or oblique orientation to the long axis of the tooth in an apical direction. In contrast, the defect is perpendicular to the long axis of the tooth in supra-alveolar bone loss. Unlike intra-alveolar bone defects, supra-alveolar bone defects lack supporting adjacent space, which makes supra-alveolar bone regeneration more challenging. In addition, the limited availability of resources in terms of vascularity and underlying tissues is another obstacle to supra-alveolar bone regeneration. Currently, supra-alveolar bone loss is the least predictable periodontal defect type in regenerative periodontal therapy. In addition, supra-alveolar bone loss is much more common than other alveolar bone loss. Despite its prevalence, research on supra-alveolar bone regeneration remains sparse, indicating an unmet need for significant research efforts in this area. This review summarize recent advances, obstacles, and future directions in the field of supra-alveolar bone regeneration. We discuss the biomaterials, bioactive molecules, and cells that have been tested for supra-alveolar bone regeneration, followed by pre-clinical and clinical approaches employed in this field. Additionally, we highlight obstacles and present future directions that will propel supra-alveolar bone research forward.

Comparative evaluation of gingival fibroblast growth on 3D-printed and milled zirconia: An in vitro study.

PURPOSE: The purpose of this study was to analyze the fibroblast growth and proliferation on 3D-printed zirconia in presence and absence of porosities. MATERIAL AND METHODS: A total of 40 bars (8 × 4 × 3) were included in this study. Thirty 3D-printed and 10 milled zirconia samples were prepared. The 3D-printed samples had different porosities, 0% (PZ0), 20% (PZ20), and 40% (PZ40) with 10 specimens in each group. Milled zirconia samples were used as the control (MZ). Rat gingival fibroblasts were cultured for 48 h, and the proliferation of fibroblasts on each sample in each group (n = 10) was determined by MTT assays. The differences among the four groups were compared by one-way ANOVA. To test the significance of the observed differences between two groups, an unpaired Student's t-test was applied. The significance level was set at p < 0.05. Qualitative analysis for the cell culture was performed using scanning electron microscopy. RESULTS: One-way ANOVA showed that the numbers of the fibroblasts among the four groups had a statistical difference. Post hoc Bonferroni test revealed that there was no significant difference between PZ0 and MZ; however, all other groups and among groups were significantly different. CONCLUSIONS: Fibroblasts had a better affinity toward the MZ and PZ0 in a short period of cell culture time.

Denervation delays initial bone healing of rat tooth extraction socket.

Functions of nerves on bone has been a subject of intense research. The aim of this study is to observe initial bone healing of rat tooth extraction socket after inferior alveolar nerve transection. The bilateral mandible second molars of eighteen Wistar rats were extracted in the study. The rats also suffered from right inferior alveolar nerve transection simultaneously (D + E group), only extraction as control (E group). One, two and four weeks after extraction, the mandibles were taken out for histological observation, TRAP staining, immunofluorescence, immunohistochemistry and micro-computed tomography (Micro-CT). Mouse bone marrow derived macrophages (BMMs) were cultured in vitro. Expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) were detected in vivo and vitro. The alveolar sockets had been filled to a large extent with new bone at 4 weeks, but BV/TV and BMD decreased in the D + E group. Accordingly, Expressions of osteocalcin (OCN) and osteopontin (OPN) were down-regulated in the D + E group. Denervation increased TRAP-positive osteoclasts and decreased expressions of Nrf2 at 2 weeks after extraction. Decreased Nrf2 promoted osteoclast differentiation of BMMs in vitro. Denervation delays initial bone healing of rat tooth extraction socket. Osteoclast activation induced by decreased Nrf2 might participated in the process.

Quantitative characterizations of the Sharpey's fibers of rat molars.

BACKGROUND AND OBJECTIVE: The Sharpey's fibers of periodontal ligament (PDL) anchor the PDL to alveolar bone and cementum and are essential for the function of PDL. While qualitative analyses of the Sharpey's fibers have been widely explored, a comprehensive quantitative characterization of the Sharpey's fibers is not available. In this work, we selected rat molars as a model and comprehensively characterized the PDL Sharpey's fibers (diameter, density, length, embedding angle, and insertion angle). MATERIALS AND METHODS: A total of 24 rat mandibular molars, eight maxillary first molars, and their surrounding alveolar bone were harvested, fixed, rendered anorganic and observed under scanning electron microscopy (SEM). The mandibles and maxillae (n = 4) were harvested, processed, sectioned, and stained with Sirius red for histological observation. SEM images were used for quantitative analyses of diameters and densities of the Sharpey's fibers, while Sirius red staining images were used to measure lengths and angles. The Sharpey's fibers were comprehensively characterized in terms of positions (cervical, middle, and apical thirds), PDL fiber groups (alveolar crest, horizontal, oblique, apical, and interradicular groups), sides (cementum and bone sides), and teeth (mandibular first, second, third molars, and maxillary first molar). RESULTS: Our results showed that the characteristic parameters of the Sharpey's fibers varied in different positions, fiber groups, sides, and teeth. Specifically, the median diameter of the Sharpey's fibers on the bone side was significantly greater than that on the cementum side, while the median density of the Sharpey's fibers on the bone side was significantly lower than that on the cementum side, regardless of the positions and teeth. For the same tooth, the median length of the embedded Sharpey's fibers on the bone side was more than two times greater than that on the cementum side. Among all fiber groups, the alveolar crest group had the maximum length of the Sharpey's fibers on the bone side and the minimal length of the Sharpey's fibers on the cementum side. There is an approximate 5-15° difference between the embedding angle and the insertion angle in each group. The oblique group had the smallest embedding angles on both the bone and cementum sides. CONCLUSION: This study provides a comprehensive and quantitative characterization of the Sharpey's fibers using rat molars as a model. Overall, these parameters varied according to different vertical positions, fiber groups, teeth, and jawbones. The quantitative information of the Sharpey's fibers presented in this work facilitates our understanding of PDL functions and advances the development of biomimetic materials for periodontal tissue regeneration.

Injectable scaffolds: Preparation and application in dental and craniofacial regeneration.

Injectable scaffolds are appealing for tissue regeneration because they offer many advantages over pre-formed scaffolds. This article provides a comprehensive review of the injectable scaffolds currently being investigated for dental and craniofacial tissue regeneration. First, we provide an overview of injectable scaffolding materials, including natural, synthetic, and composite biomaterials. Next, we discuss a variety of characteristic parameters and gelation mechanisms of the injectable scaffolds. The advanced injectable scaffolding systems developed in recent years are then illustrated. Furthermore, we summarize the applications of the injectable scaffolds for the regeneration of dental and craniofacial tissues that include pulp, dentin, periodontal ligament, temporomandibular joint, and alveolar bone. Finally, our perspectives on the injectable scaffolds for dental and craniofacial tissue regeneration are offered as signposts for the future advancement of this field.

Multifunctional Nanofibrous Hollow Microspheres for Enhanced Periodontal Bone Regeneration.

Destructive periodontitis destroys alveolar bone and eventually leads to tooth loss. While guided bone regeneration, which is based on creating a physical barrier to hinder the infiltration of epithelial and connective tissues into defect sites, has been widely used for alveolar bone regeneration, its outcomes remain variable. In this work, a multifunctional nanofibrous hollow microsphere (NFHMS) is developed for enhanced alveolar bone regeneration. The NFHMS is first prepared via combining a double emulsification and a thermally induced phase separation process. Next, E7, a short peptide with high specific affinity to bone marrow-derived stem cells (BMSCs), is conjugated onto the surface of NFHMS. After that, bone forming peptide (BFP), a short peptide derived from bone morphology protein 7 is loaded in calcium phosphate (CaP) nanoparticles, which are further encapsulated in the hollow space of the NFHMS-E7 to form NFHMS-E7-CaP/BFP. The NFHMS-E7-CaP/BFP selectively promoted the adhesion of BMSCs and expelled the adhesion of fibroblasts and epithelial cells. In addition, the BFP is sustainedly released from the NFHMS-E7-CaP/BFP to enhance the osteogenesis of BMSCs. A rat challenging fenestration defect model showed that the NFHMS-E7-CaP/BFP significantly enhanced alveolar bone tissue regeneration. This work provides a novel bioengineering approach for guided bone regeneration.

Effects of PEG antibodies on in vivo performance of LNP-mRNA vaccines.

Polyethylene glycol (PEG) plays important roles in stabilizing and lengthening circulation time of lipid nanoparticle (LNP) vaccines. Nowadays various levels of PEG antibodies have been detected in human blood, but the impact and mechanism of PEG antibodies on the in vivo performance of LNP vaccines has not been clarified thoroughly. By illustrating the distribution characteristics of PEG antibodies in human, the present study focused on the influence of PEG antibodies on the safety and efficacy of LNP-mRNA vaccine against COVID-19 in animal models. It was found that PEG antibodies led to shortened blood circulation duration, elevated accumulation and mRNA expression in liver and spleen, enhanced expression in macrophage and dendritic cells, while without affecting the production of anti-Spike protein antibodies of COVID-19 LNP vaccine. Noteworthily, PEG antibodies binding on the LNP vaccine increased probability of complement activation in animal as well as in human serum and led to lethal side effect in large dosage via intravenous injection of mice. Our data suggested that PEG antibodies in human was a risky factor of LNP-based vaccines for biosafety concerns but not efficacy.

Efficacy of Different Solvents for Achieving Patency in Teeth Obturated Using Bioceramic Sealer.

INTRODUCTION: The retreatment of teeth obturated with bioceramic sealers has been shown to be challenging. The purpose of this study was to compare the efficacy of 10% formic acid (FA), 20% hydrochloric acid (HCl), and chloroform for achieving patency in teeth obturated with a bioceramic sealer. METHODS: Forty-five extracted single-canal teeth were instrumented and obturated 1.5 mm short of the working length using gutta-percha and EndoSequence BC Sealer (Brasseler USA, Savannah, GA). Two weeks later, gutta-percha was removed, and apical patency was attempted with a 10 C-file and a solvent (FA, HCl, or chloroform). The time to gain patency through EndoSequence BC Sealer was recorded. Five additional teeth were irrigated with the solvents, and scanning electron microscopy was used to evaluate the effect of solvents on the dentin. The Kruskal-Wallis and Mann-Whitney tests were used for statistical analysis. RESULTS: Patency was achieved in all canals, except for 1 in the chloroform group. Regarding the time to achieve patency, no significant difference was noted between chloroform and FA (P > .05). However, there was a significant difference between chloroform and HCl (P < .05) and between FA and HCl (P < .05). The erosion of dentinal tubules was more evident when HCl was used compared with the other irrigants. CONCLUSIONS: Twenty percent HCl was superior to 10% FA and chloroform in achieving patency in teeth obturated with EndoSequence BC Sealer. Regardless of the solvent used, patency may be achieved for most of the cases obturated with gutta-percha and EndoSequence BC Sealer.

E7-Conjugated Bio-Inspired Microspheres as a Biological Barrier for Guided Tissue Regeneration.

Guided tissue regeneration (GTR), which is based on creating a physical barrier to prevent the downgrowth of epithelial and connective tissues into the defect site, has been widely used in clinical practice for periodontal regeneration for many years. However, its outcomes remain variable due to highly specific indications, the demand for proficient surgical skills, and frequent occurrence of complications. In this study, we developed a new GTR biomaterial that acts as a biological barrier for epithelial cells and fibroblasts while also serving as a scaffold for bone marrow-derived mesenchymal stem cells (BMSCs) and periodontal ligament stem cells (PDLSCs). This innovative GTR biomaterial is bioinspired injectable microspheres that are self-assembled from nanofibers, and their surfaces are conjugated with E7, a short peptide that selectively promotes BMSC and PDLSC adhesion but inhibits the attachment and spreading of epithelial cells and gingival fibroblasts. The selective affinity afforded by E7 on the surfaces of the nanofibrous microspheres facilitated the colonization of BMSCs in the periodontal defect, thereby substantially improving functional periodontal regeneration, as evidenced by enhanced new bone formation, reduced root exposure, and diminished attachment loss. The remarkable superiority of the bioinspired microspheres over conventional GTR materials in promoting periodontal regeneration underscores the potential of this innovative approach to enhance the efficacy of functional periodontal tissue regeneration.

Functional microspheres for tissue regeneration.

As a new type of injectable biomaterials, functional microspheres have attracted increasing attention in tissue regeneration because they possess some advantageous properties compared to other biomaterials, including hydrogels. A variety of bio-inspired microspheres with unique structures and properties have been developed as cellular carriers and drug delivery vehicles in recent years. In this review, we provide a comprehensive summary of the progress of functional and biodegradable microspheres that have been used for tissue regeneration over the last two decades. First, we briefly introduce the biomaterials and general methods for microsphere fabrication. Next, we focus on the newly developed technologies for preparing functional microspheres, including macroporous microspheres, nanofibrous microspheres, hollow microspheres, core-shell structured microspheres, and surface-modified functional microspheres. After that, we discuss the application of functional microspheres for tissue regeneration, specifically for bone, cartilage, dental, neural, cardiac, and skin tissue regeneration. Last, we present our perspectives and future directions of functional microspheres as injectable carriers for the future advancement of tissue regeneration.

TIPE2 regulates periodontal inflammation by inhibiting NF-κB p65 phosphorylation.

BACKGROUND: The roles and molecular mechanisms of tumor necrosis factor-α-induced protein 8-like 2 (TIPE2) in periodontitis remain largely unknown. OBJECTIVE: This study aimed to determine the expression of TIPE2 and NF-κB p65 in rat Porphyromonas gingivalis-induced periodontics in vivo. METHODOLOGY: Periodontal inflammation and alveolar bone resorption were analyzed using western blotting, micro-computed tomography, TRAP staining, immunohistochemistry, and immunofluorescence. THP-1 monocytes were stimulated using 1 µg/ml Pg. lipopolysaccharide (Pg.LPS) to determine the expression of TIPE2 in vitro. TIPE2 mRNA was suppressed by siRNA transfection, and the transfection efficiency was proven using western blotting and real-time PCR. The NF-κB pathway was activated by treating the cells with 1 µg/ml Pg.LPS to explore related mechanisms. RESULTS: The expression of both TIPE2 and NF-κB p65 was increased in the gingival tissues of rat periodontitis compared with normal tissues. Positive expression of TIPE2 was distributed in inflammatory infiltrating cells and osteoclasts in the marginal lacunae of the alveolar bone. However, strong positive expression of TIPE2 in THP-1 was downregulated after Pg.LPS stimulation. TIPE2 levels negatively correlated with TNF-α and IL-1ß. Decreased TIPE2 in THP-1 further promoted NF-κB p65 phosphorylation. Mechanistically, TIPE2 knockdown upregulated NF-κB signaling pathway activity. CONCLUSIONS: Taken together, these findings demonstrate that TIPE2 knockdown aggravates periodontal inflammatory infiltration via NF-κB pathway. Interventions aimed at increasing TIPE2 may help in the therapeutic applications for periodontitis.

Nanofibrous hollow microspheres self-assembled from star-shaped polymers as injectable cell carriers for knee repair.

To repair complexly shaped tissue defects, an injectable cell carrier is desirable to achieve an accurate fit and to minimize surgical intervention. However, the injectable carriers available at present have limitations, and are not used clinically for cartilage regeneration. Here, we report nanofibrous hollow microspheres self-assembled from star-shaped biodegradable polymers as an injectable cell carrier. The nanofibrous hollow microspheres, integrating the extracellular-matrix-mimicking architecture with a highly porous injectable form, were shown to efficiently accommodate cells and enhance cartilage regeneration, compared with control microspheres. The nanofibrous hollow microspheres also supported a significantly larger amount of, and higher-quality, cartilage regeneration than the chondrocytes-alone group in an ectopic implantation model. In a critical-size rabbit osteochondral defect-repair model, the nanofibrous hollow microspheres/chondrocytes group achieved substantially better cartilage repair than the chondrocytes-alone group that simulates the clinically available autologous chondrocyte implantation procedure. These results indicate that the nanofibrous hollow microspheres are an excellent injectable cell carrier for cartilage regeneration.

Biomaterials for Periodontal Regeneration.

As a widespread chronical disease, periodontitis progressively destroys tooth-supporting structures (periodontium) and eventually leads to tooth loss. Therefore, regeneration of damaged/lost periodontal tissues has been a major subject in periodontal research. During periodontal tissue regeneration, biomaterials play pivotal roles in improving the outcome of the periodontal therapy. With the advancement of biomaterial science and engineering in recent years, new biomimetic materials and scaffolding fabrication technologies have been proposed for periodontal tissue regeneration. This article summarizes recent progress in periodontal tissue regeneration from a biomaterial perspective. First, various guide tissue regeneration/guide bone regeneration membranes and grafting biomaterials for periodontal tissue regeneration are overviewed. Next, the recent development of multifunctional scaffolding biomaterials for alveolar bone/periodontal ligament/cementum regeneration is summarized. Finally, clinical care points and perspectives on the use of biomimetic scaffolding materials to reconstruct the hierarchical periodontal tissues are provided.

Delivering Multifunctional Peptide-Conjugated Gene Carrier/miRNA-218 Complexes from Monodisperse Microspheres for Bone Regeneration.

MicroRNAs (miRNAs) play a pivotal role in regulating gene expression and are considered new molecular targets in bone tissue engineering. However, effective delivery of miRNAs to the defect areas and transfection of the miRNAs into osteogenic progenitor cells has been an obstacle in the application. In this work, miRNA-218 (miR-218) was used as an osteogenic miRNA regulator, and a multifunctional peptide-conjugated gene carrier poly(lactide-co-glycolide)-g-polyethylenimine-b-polyethylene glycol-R9-G4-IKVAVW (PPP-RGI) was developed to condense with miR-218 to form PPP-RGI/miR-218 complexes that were further encapsulated into monodisperse injectable microspheres for enhanced bone regeneration. The PPP-RGI was synthesized via conjugating R9-G4-IKVAVW (RGI), a multifunctional peptide, onto poly(lactide-co-glycolide)-g-polyethylenimine-b-polyethylene glycol (PPP). A microfluidic and synchronous photo-cross-linking process was further developed to encapsulate the PPP-RGI/miR-218 complexes into monodisperse gelatin methacryloyl microspheres. The monodisperse microspheres controlled the delivery of PPP-RGI/miR-218 to the designated defect site, and PPP-RGI facilitated the transfection of miR-218 into osteogenic progenitor cells. An in vivo calvarial defect model showed that the PPP-RGI/miR-218-loaded microspheres significantly enhanced bone tissue regeneration. This work provides a novel approach to effectively deliver miRNA and transfect targeting cells in vivo for advanced regenerative therapies.

Biomimetic Tubular Matrix Induces Periodontal Ligament Principal Fiber Formation and Inhibits Osteogenic Differentiation of Periodontal Ligament Stem Cells.

Periodontal ligament (PDL) is assembled from highly organized collagen fiber bundles (PDL principal fibers) that are crucial in supporting teeth and buffering mechanical force. Therefore, regeneration of PDL needs to reconstruct these well-ordered fiber bundles to restore PDL functions. However, the formation of PDL principal fibers has long been a challenge due to the absence of an effective three-dimensional (3D) matrix to guide the growth of periodontal ligament stem cells (PDLSCs) and to inhibit the osteogenic differentiation of PDLSCs during the PDL principal fibers deposition. In this work, we designed and fabricated a bio-inspired tubular 3D matrix to guide the migration and growth of human PDLSCs and form well-aligned PDL principal fibers. As a biomimetic 3D template, the tubular matrix controlled PDLSCs migration inside the tubules and aligned the cells to the designated direction. Inside the tubular matrix, the PDLSCs expressed PDL markers and formed oriented fiber bundles with the same size and density as those of natural PDL principal fibers. Furthermore, the tubular matrix downregulated the osteogenic differentiation of PDLSCs. A mechanism study revealed that the Yap1/Twist1 signaling pathway was involved in the inhibition of PDLSCs osteogenesis within the tubular matrix. This work provides an effective approach to induce PDLSCs to form principal fibers and gives insight into the underlying mechanism of inhibiting the osteogenic differentiation of PDLSCs in biomimetic tubular matrices.

Guar Gum/Ethyl Cellulose-Polyvinyl Pyrrolidone Composite-Based Quartz Crystal Microbalance Humidity Sensor for Human Respiration Monitoring.

In this work, the guar gum (GG) and the electrospinned ethyl cellulose-polyvinyl pyrrolidone (EC-PVP) nanofibers were used as humidity-sensitive materials for fabricating a quartz crystal microbalance (QCM) sensor. Fourier transform infrared spectroscopy, scanning electron microscopy, water contact angle test, and X-ray photoelectron spectra were used to characterize the synthesized GG/EC-PVP composite material, confirming its successful preparation and good hydrophilicity. The humidity sensitivity experiments were performed at room temperature. The GG/EC-PVP-coated QCM sensor has high sensitivity (55.72 Hz/%RH) and low hysteresis (2.8% RH) in a wide relative humidity range (0-97% RH), short response/recovery time (26/2 s), excellent selectivity, good repeatability, and stability. The combined action of hydrophilic groups and porous structure enhances the humidity sensitivity. The GG/EC-PVP sensor can be used to capture and measure typical breathing patterns in different human basic emotions due to its good performance. Furthermore, a lie-detector system was also designed for judging the lying through detecting the emotional breathing pattern of the subjects.

Long-acting PFI-2 small molecule release and multilayer scaffold design achieve extensive new formation of complex periodontal tissues with unprecedented fidelity.

The faithful engineering of complex human tissues such as the bone/soft tissue/mineralized tissue interface in periodontal tissues requires innovative molecular cues in conjunction with tailored scaffolds. To address the loss of periodontal bone and connective tissues following periodontal disease, we have generated a polydopamine and collagen coated electrospun PLGA-PCL (PP) scaffold enriched with the small molecule mediator PFI-2 (PP-PFI-pDA-COL-PFI). In vitro 3D studies using PDL progenitors revealed that the PP-PFI-pDA-COL-PFI scaffold substantially enhanced Alizarin Red staining, increased Ca/P ratios 4-fold, and stimulated cell proliferation more than 12-fold compared to PP-controls, suggestive of its potential for mineralized tissue engineering. When applied in our experimental periodontitis model, the PP-PFI-pDA-COL-PFI scaffold resulted in a substantial 34% reduction in alveolar bone defect height, a 25% root-length gain in periodontal attachment, and the formation of highly ordered regenerated acellular cementum twice as thick as in controls. Explaining the mechanism of PFI-2 mineralized tissue regeneration in periodontal tissues, PFI-2 inhibited SETD7-mediated ß-Catenin protein methylation and increased ß-Catenin nuclear localization. Together, dual-level PFI-2 incorporation into a degradable, dopamine/collagen coated PLGA/PCL scaffold backbone resulted in the regeneration of the tripartite periodontal complex with unprecedented fidelity, including periodontal attachment and new formation of mineralized tissues in inflamed periodontal environments.

Surface Finishing of FDM-Fabricated Amorphous Polyetheretherketone and Its Carbon-Fiber-Reinforced Composite by Dry Milling.

In recent years, many investigations have been devoted to fused deposition modeling (FDM) of high-performance polymer-polyetheretherketone (PEEK) and carbon-fiber-reinforced PEEK (CF/PEEK) for biomedical and aerospace applications. However, the staircase effect naturally brought about by FDM restricts further applications of 3D-printed PEEK and its composites in high-temperature molds, medical implants, and precision components, which require better or customized surface qualities. Hence, this work aimed to reduce the staircase effect and improve the surface quality of 3D-printed PEEK and CF/PEEK parts by dry milling of the fluctuant exterior surface. The co-dependency between 3D printing parameters (raster angle and layer thickness) and milling parameters (depth of cut, spindle speed, and feed rate per tooth) were investigated through experiments. The difference in removal mechanisms for PEEK and CF/PEEK was revealed. It was confirmed that the smearing effect enhanced the surface quality based on the morphology analysis and the simulation model. Both the raster angle of +45°/-45° and the small layer thickness could improve the surface quality of these 3D-printed polymers after dry milling. A large depth of cut and a large feed rate per tooth were likely to deteriorate the finished polymer surface. The spindle speed could influence the morphologies without significant changes in roughness values. Finally, a demonstration was performed to verify that dry milling of 3D-printed amorphous PEEK and CF/PEEK parts could lead to a high surface quality for critical requirements.

Multifunctional peptide-conjugated nanocarriers for pulp regeneration in a full-length human tooth root.

Dental pulp is a highly vascularized tissue, situated in an inextensible environment surrounded by rigid dentinal walls. The pulp receives its blood supply solely from the small apical foramen of a tooth root. Due to the unique anatomy that controls nutrition supply, regeneration of pulp tissue in a full-length tooth root has long been a challenge in regenerative endodontics. In this study, we designed and synthesized a multifunctional peptide-conjugated, pH-sensitive, non-viral gene vector for fast revascularization and pulp regeneration in a full-length human tooth root. The multifunctional peptide was designed to have distinctive features, including a cell-penetrating peptide to enhance cellular uptake, a nuclear localization signal peptide to assist in the translocation of an angiogenic gene into the nucleus, and a fluorescent tryptophan residue to visualize and quantify the transfection efficiency. Furthermore, a pH-sensitive dimethylmaleic anhydride (DMA) was integrated with the multifunctional peptide to enhance the transfected gene complex to escape from endosomes/lysosomes after internalization. In vitro experiments showed that the multifunctional non-viral gene vector significantly increased internalization and gene transfection efficiency as well as reduced cytotoxicity. After dental pulp stem cells (DPSCs) were transfected with the multifunctional gene vector/pVEGF complexes, the expression of VEGF from the DPSCs was upregulated for more than eight folds, which in turn greatly enhanced endothelial cell migration and vascular-like tube formation. Six weeks after implantation, the VEGF-transfected DPSCs accelerated new blood vessel formation and the regenerated pulp tissue occupied most of the area in the canal of a full-length human tooth root. The multifunctional peptide conjugated non-viral gene delivery is a safe and effective approach for regenerative endodontics. STATEMENT OF SIGNIFICANCE: Pulp regeneration in a full-length tooth root canal has long been a challenge in regenerative endodontics. This is due to the unique root anatomy that allows the blood supply of the tooth root only from a small apical foramen (< 1 mm), leading to a severe barrier for revascularization during pulp regeneration. In this work, we designed a multifunctional peptide-conjugated, pH-sensitive, non-viral gene vector to address this challenge. Our work shows that the peptide-conjugated system was an excellent carrier for fast revascularization and pulp tissue regeneration in a full-length toot root. This study will interest the multidisciplinary readership in gene delivery, biomaterials, and dental/craniofacial tissue engineering community.